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Competition-Mediated FRET-Switching DNA Tetrahedron Molecular Beacon for Intracellular Molecular Detection
journal contribution
posted on 2016-12-05, 00:00 authored by Nuli Xie, Jin Huang, Xiaohai Yang, Yanjing Yang, Ke Quan, Min Ou, Hongmei Fang, Kemin WangRecently, DNA tetrahedron-based sensors
for intracellular detection
have attracted more attention due to many interesting properties,
including good structural rigidity, excellent biocompatibility, high
resistance to enzymatic degradation, and the ability to enter cells
without the use of transfection agents. However, the previous designs
are still restricted by their lack of accuracy, reliability, and generality.
Herein, to solve these limitations, we describe self-assembly of the
competition-mediated FRET-switching DNA tetrahedron molecular beacon
(CF-DTMB), and its applications for intracellular tumor-related mRNA
detection. In brief, the recognition strand is partially complementary
to its competitor, which is a hairpin stem-loop structure inserted
in one edge of the DNA tetrahedron. In the absence of a target, a
long domain of the recognition strand hybridizes with the competitor
and makes the hairpin structure open, inducing the two labeled dyes
to be spatially separated (FRET off). However, in the presence of
the target, the competitor is substituted for the target to bind with
the recognition strand, subsequently leading the formation of a stem-loop
structure, which draws two dyes together (FRET on). The results demonstrate
that the current strategy possesses the merits of the previous DNA
tetrahedron-based sensors, but also improves the accuracy and reliability.
Furthermore, we have also demonstrated that our design can become
a general strategy for detecting and imaging a variety of other molecules,
such as adenosine triphosphate (ATP) in living cells. Therefore, the
CF-DTMB can serve as an excellent intracellular molecular detection
tool, which is promising for biological and disease studies.
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CF-DTMBIntracellular Molecular DetectionreliabilityATPcompetition-mediated FRET-switching DNA tetrahedrondyeCompetition-Mediated FRET-Switching DNA Tetrahedron Molecular Beaconrecognition strand hybridizesaccuracyintracellular tumor-related mRNA detectionDNA tetrahedron-based sensorsstrategyhairpin stem-loop structurerecognition strand
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